The mission of the Molecular Core is to facilitate the introduction and use of molecular genetic methods by a range of potential DERC investigators at Yale. The Core's central goal is to provide: a) services, b) training, c) consultation and d) equipment and resources dedicated to support a number of essential molecular techniques. The functions of the Molecular Core are now centered on two principal services: 1) the construction of transgenes and gene knock-out vectors, and 2) in situ hybridization of cells and tissues. A third major area of need has recently been identified relating to quantitation of mRNA expression levels by real-time PCR. In the next grant cycle, we therefore intend to offer expanded services in this area. Additional services frequendy utilized by DERC investigators include: genotyping of transgenic, knockout and naturally occurring mutant mice, site-directed mutagenesis, primer design for a variety of PCR-related applications, RNase protection assays and construction of riboprobes. Advice and consultation are offered for a variety of related methodologies, including Southern and Northern blotting, eDNA and genomic library construction, reporter gene assays, and PCR-based techniques.